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https://www.readbyqxmd.com/read/27924363/characteristics-and-applications-of-recombinant-thermostable-amylopullulanase-of-geobacillus-thermoleovorans-secreted-by-pichia-pastoris
#1
M Nisha, T Satyanarayana
The 3'-deleted amylopullulanase gene from the extreme thermophile Geobacillus thermoleovorans (Gt-apuΔC) was expressed extracellularly in Pichia pastoris under both methanol-inducible AOX1 and constitutive GAP promoters. The expression of the gene (Gt-apuΔC) was higher under GAP promoter (36.2 U ml(-1), α-amylase; 33.5 U ml(-1), pullulanase) than that under AOX1 promoter (32.5 and 28.6 U ml(-1)). The heavily glycosylated Gt-apuΔC from the recombinant P. pastoris displays higher substrate specificity, thermal stability and starch saccharification efficiency than that expressed in Escherichia coli...
December 6, 2016: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/27921412/impacts-of-thermal-treatments-on-major-and-minor-allergens-of-sea-snail-cerithidea-obtusa-obtuse-horn-shell
#2
Rosmilah Misnan, Norazlin Salahudin Abd Aziz, Zailatul Hani Mohamad Yadzir, Faizal Bakhtiar, Noormalin Abdullah, Shahnaz Murad
Snail is one of the worst causes of food allergy. Thus, the aim of this study was to identify the major and minor allergens of the local marine snail (Cerithidea obtusa) and subsequently to investigate the impacts of heat treatment on the IgE-binding activity of snail allergens. Proteins from raw and heat-treated snails (boiled, roasted and fried) were extracted and then resolved by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Immunoblotting of all extracts were then performed using sera from patients with snail allergy...
August 2016: Iranian Journal of Allergy, Asthma, and Immunology
https://www.readbyqxmd.com/read/27920296/a-sulfur-oxygenase-from-the-haloalkaliphilic-bacterium-thioalkalivibrio-paradoxus-with-atypically-low-reductase-activity
#3
Patrick Rühl, Uwe Pöll, Johannes Braun, Andreas Klingl, Arnulf Kletzin
: Sequence comparisons showed that the sulfur oxygenase reductase (SOR) of the haloalkaliphilic bacterium Thioalkalivibrio paradoxus Arh 1 (TpSOR) is branching deeply within dendrograms of these proteins (29-34% identity). A synthetic gene encoding the TpSOR expressed in E. coli resulted in a protein of 14.7±0.9 nm in diameter and an apparent molecular mass of 556 kDa. Sulfite and thiosulfate were formed from elemental sulfur in a temperature range of 10-98°C (Topt≈80°C) and a pH range of 6-11...
December 5, 2016: Journal of Bacteriology
https://www.readbyqxmd.com/read/27917828/molecular-determinants-of-substrate-specificity-revealed-by-the-structure-of-clostridium-thermocellum-arabinofuranosidase-43a-from-glycosyl-hydrolase-family-43-subfamily-16
#4
Arun Goyal, Shadab Ahmed, Kedar Sharma, Vikas Gupta, Pedro Bule, Victor D Alves, Carlos M G A Fontes, Shabir Najmudin
The recent division of the large glycoside hydrolase family 43 (GH43) into subfamilies offers a renewed opportunity to develop structure-function studies aimed at clarifying the molecular determinants of substrate specificity in carbohydrate-degrading enzymes. α-L-Arabinofuranosidases (EC 3.2.1.55) remove arabinose side chains from heteropolysaccharides such as xylan and arabinan. However, there is some evidence suggesting that arabinofuranosidases are substrate-specific, being unable to display a debranching activity on different polysaccharides...
December 1, 2016: Acta Crystallographica. Section D, Structural Biology
https://www.readbyqxmd.com/read/27917251/droplet-millifluidics-for-kinetic-study-of-transketolase
#5
A Pinsolle, F Charmantray, L Hecquet, F Sarrazin
We present a continuous-flow reactor at the millifluidic scale coupled with an online, non-intrusive spectroscopic monitoring method for determining the kinetic parameters of an enzyme, transketolase (TK) used in biocatalysis for the synthesis of polyols by carboligation. The millifluidic system used is based on droplet flow, a well-established method for kinetic chemical data acquisition. The TK assay is based on the direct quantitative measurement of bicarbonate ions released during the transketolase-catalysed reaction in the presence of hydroxypyruvic acid as the donor, thanks to an irreversible reaction: bicarbonate ions react with phosphoenolpyruvate (PEP) in the presence of PEP carboxylase as the first auxiliary enzyme...
November 2016: Biomicrofluidics
https://www.readbyqxmd.com/read/27915377/crystal-structure-and-characterization-of-esterase-est25-mutants-reveal-improved-enantioselectivity-toward-s-ketoprofen-ethyl-ester
#6
Jinyeong Kim, Seung-Hyeon Seok, Eunsoo Hong, Tae Hyeon Yoo, Min-Duk Seo, Yeonwoo Ryu
Esterases comprise a group of enzymes that catalyze the cleavage and synthesis of ester bonds. They are important in biotechnological applications owing to their enantioselectivity, regioselectivity, broad substrate specificity, and the fact that they do not require cofactors. In a previous study, we isolated the esterase Est25 from a metagenomic library. Est25 showed catalytic activity toward the (R,S)-ketoprofen ethyl ester but had low enantioselectivity toward the (S)-ketoprofen ethyl ester. Because (S)-ketoprofen has stronger anti-inflammatory effects and fewer side effects than (R)-ketoprofen, enantioselectivity of this esterase is important...
December 3, 2016: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/27914120/effect-of-glycosylation-on-the-biocatalytic-properties-of-hydroxynitrile-lyase-from-the-passion-fruit-passiflora-edulis-a-comparison-of-natural-and-recombinant-enzymes
#7
Aem Nuylert, Yuko Ishida, Yasuhisa Asano
A hydroxynitrile lyase from the passion fruit, Passiflora edulis (PeHNL) was isolated from the leaves and showed high stability in biphasic co-organic solvent systems for cyanohydrin synthesis. Cyanohydrins are important building blocks for the production of fine chemicals and pharmaceuticals. Thus, to enhance production yields of PeHNL for industrial applications, we cloned and expressed recombinant PeHNL in E. coli BL21 (DE3) and P. pastoris GS115 cells without a signal peptide sequence. The aim of the present study is to determine the effect of N-glycosylation on enzyme stability and catalytic properties in microbial expression systems...
December 3, 2016: Chembiochem: a European Journal of Chemical Biology
https://www.readbyqxmd.com/read/27909846/benzimidazole-inhibitors-of-protein-kinase-ck2-potently-inhibit-the-activity-of-atypical-protein-kinase-rio1
#8
Konrad Kubiński, Maciej Masłyk, Andrzej Orzeszko
Benzimidazole derivatives of 5,6-dichlorobenzimidazole 1-β-D-ribofuranoside (DRB) comprise the important class of protein kinase CK2 inhibitors. Depending on the structure, benzimidazoles inhibit CK2 with different selectivity and potency. Besides CK2, the compounds can inhibit, with similar activity, other classical eukaryotic protein kinases (e.g. PIM, DYRK, and PKD). The present results show that a majority of the most common CK2 inhibitors can affect the atypical kinase Rio1 in a nanomolar range. Kinetic data confirmed the mode of action of benzimidazoles as typical ATP-competitive inhibitors...
December 1, 2016: Molecular and Cellular Biochemistry
https://www.readbyqxmd.com/read/27909291/engineering-of-a-thermostable-esterase-est816-to-improve-its-quorum-quenching-activity-and-the-underlying-structural-basis
#9
Xiwen Liu, Li-Chuang Cao, Xin-Jiong Fan, Yu-Huan Liu, Wei Xie
N-acyl-homoserine lactones (AHLs) are small diffusible molecules called autoinducers that mediate cell-to-cell communications. Enzymatic degradation of AHLs is a promising bio-control strategy known as quorum-quenching. To improve the quorum-quenching activity of a thermostable esterase Est816, which had been previously cloned, we have engineered the enzyme by random mutagenesis. One of the mutants M2 with double amino acid substitutions (A216V/K238N) showed 3-fold improvement on catalytic efficiency. Based on the crystal structure determined at 2...
December 2, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27904395/purification-and-characterization-of-thermostable-and-detergent-stable-%C3%AE-amylase-from-anoxybacillus-sp-ah1
#10
Ömer Acer, Fatma Matpan Bekler, Hemşe Pirinççioğlu, Reyhan Gül Güven, Kemal Güven
A thermostable and detergent-stable α-amylase from a newly isolated Anoxybacillus sp. AH1 was purified and characterized. Maximum enzyme production (1874.8 U/mL) was obtained at 24 h of incubation. The amylase was purified by using Sephadex G-75 gel filtration, after which an 18-fold increase in specific activity and a yield of 9% were achieved. The molecular mass of the purified enzyme was estimated at 85 kDa by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimum pH and temperature values of the enzyme were 7...
March 2016: Food Technology and Biotechnology
https://www.readbyqxmd.com/read/27902339/progressive-glycosylation-of-the-hemagglutinin-of-avian-influenza-h5n1-modulates-virus-replication-virulence-and-chicken-to-chicken-transmission-without-significant-impact-on-antigenic-drift
#11
El-Sayed M Abdelwhab, Jutta Veits, Kerstin Tauscher, Mario Ziller, Christian Grund, Mohamed K Hassan, Momtaz Shaheen, Timm C Harder, Jens P Teifke, Jürgen Stech, Thomas C Mettenleiter
Highly pathogenic H5N1 avian influenza virus (A/H5N1) devastated the poultry industry and continues to pose a pandemic threat. Studying the progressive genetic changes in A/H5N1 after long term circulation in poultry may help to better understand A/H5N1 biology in birds. A/H5N1 clade 2.2.1.1 antigenic drift viruses have been isolated from vaccinated commercial poultry in Egypt. They exhibit a peculiar stepwise accumulation of glycosylation sites (GS) in the hemagglutinin (HA) with viruses carrying, beyond the conserved 5 GS, additional GS at amino acid residues 72, 154, 236 and 273 resulting in 6, 7, 8 or 9 GS in the HA...
October 28, 2016: Journal of General Virology
https://www.readbyqxmd.com/read/27900666/improved-production-of-thermostable-cellulase-from-thermoascus-aurantiacus-rckk-by-fermentation-bioprocessing-and-its-application-in-the-hydrolysis-of-office-waste-paper-algal-pulp-and-biologically-treated-wheat-straw
#12
Kavish Kumar Jain, Sandeep Kumar, Deepa Deswal, Ramesh Chander Kuhad
Thermostable cellulases have wide variety of applications and distinctive advantages, but their low titer becomes the hurdle in their commercialization. In the present work, an assessment of optimum levels of significant factors (temperature, moisture ratio, inoculum size, and ammonium sulfate) and the effect of their interactions on production of thermostable CMCase, FPase, and β-glucosidase by Thermoascus aurantiacus RCKK under solid-state fermentation (SSF) was carried out using central composite design (CCD) of response surface methodology (RSM)...
November 30, 2016: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/27900528/the-thermophilic-biomass-degrading-fungus-thielavia-terrestris-co3bag1-produces-a-hyperthermophilic-and-thermostable-%C3%AE-1-4-xylanase-with-exo-and-endo-activity
#13
Yolanda García-Huante, Maribel Cayetano-Cruz, Alejandro Santiago-Hernández, Claudia Cano-Ramírez, Rodolfo Marsch-Moreno, Jorge E Campos, Guillermo Aguilar-Osorio, Claudia G Benitez-Cardoza, Sergio Trejo-Estrada, María Eugenia Hidalgo-Lara
A hyperthermophilic and thermostable xylanase of 82 kDa (TtXynA) was purified from the culture supernatant of T. terrestris Co3Bag1, grown on carboxymethyl cellulose (CMC), and characterized biochemically. TtXynA showed optimal xylanolytic activity at pH 5.5 and at 85 °C, and retained more than 90% of its activity at a broad pH range (4.5-10). The enzyme is highly thermostable with a half-life of 23.1 days at 65 °C, and active in the presence of several metal ions. Circular dichroism spectra strongly suggest the enzyme gains secondary structures when temperature increases...
November 29, 2016: Extremophiles: Life Under Extreme Conditions
https://www.readbyqxmd.com/read/27899916/estdz3-a-new-esterolytic-enzyme-exhibiting-remarkable-thermostability
#14
Dimitra Zarafeta, Zalan Szabo, Danai Moschidi, Hien Phan, Evangelia D Chrysina, Xu Peng, Colin J Ingham, Fragiskos N Kolisis, Georgios Skretas
Lipolytic enzymes that retain high levels of catalytic activity when exposed to a variety of denaturing conditions are of high importance for a number of biotechnological applications. In this study, we aimed to identify new lipolytic enzymes, which are highly resistant to prolonged exposure to elevated temperatures. To achieve this, we searched for genes encoding for such proteins in the genomes of a microbial consortium residing in a hot spring located in China. After performing functional genomic screening on a bacterium of the genus Dictyoglomus, which was isolated from this hot spring following in situ enrichment, we identified a new esterolytic enzyme, termed EstDZ3...
2016: Frontiers in Microbiology
https://www.readbyqxmd.com/read/27899597/thermotoga-maritima-nusg-domain-interaction-mediates-autoinhibition-and-thermostability
#15
Johanna Drögemüller, Christin Schneider, Kristian Schweimer, Martin Strauß, Birgitta M Wöhrl, Paul Rösch, Stefan H Knauer
NusG, the only universally conserved transcription factor, comprises an N- and a C-terminal domain (NTD, CTD) that are flexibly connected and move independently in Escherichia coli and other organisms. In NusG from the hyperthermophilic bacterium Thermotoga maritima (tmNusG), however, NTD and CTD interact tightly. This closed state stabilizes the CTD, but masks the binding sites for the interaction partners Rho, NusE and RNA polymerase (RNAP), suggesting that tmNusG is autoinhibited. Furthermore, tmNusG and some other bacterial NusGs have an additional domain, DII, of unknown function...
November 29, 2016: Nucleic Acids Research
https://www.readbyqxmd.com/read/27899142/vancomycin-microspheres-reduce-postoperative-spine-infection-in-an-in-vivo-rabbit-model
#16
Gang Liu, Si Chen, Jun Fang, Baoshan Xu, Shuang Li, Yonghong Hao, Naif A Al-Dhabi, Shucai Deng, Veeramuthu Duraipandiyan
BACKGROUND: Surgical site infections are common and devastating complications after implants related surgeries. Staphylococcus aureus contamination is a leading cause of surgical site infections. This study aims at assessing the effect of vancomycin microspheres on reducing Staphylococcus aureus infection in an in vivo rabbit model. METHODS: Sixty surgical sites of 20 New Zealand White rabbits underwent spinal implant were randomly divided to three groups: the control group, the vancomycin group and vancomycin microspheres group...
November 29, 2016: BMC Pharmacology & Toxicology
https://www.readbyqxmd.com/read/27894826/artemin-protects-cells-and-proteins-against-oxidative-and-salt-stress
#17
Zeinab Takalloo, Reza H Sajedi, Saman Hosseinkhani, Taghi Moazzenzade
Artemin is an abundant thermostable protein in Artemia encysted embryos under environmental stresses. It is confirmed that high regulatory expression of artemin is relevant to stress resistance in this crustacean. Here, the protective role of artemin from Artemia urmiana has been investigated on survival of bacterial cells under salt and oxidative shocks. Also, for continuous monitoring of the effect of artemin in prevention of proteins aggregation/inactivation, co-expression of artemin and luciferase (as an intracellular reporter) in bacterial cells was performed...
November 25, 2016: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/27894344/siropins-novel-serine-protease-inhibitors-from-gut-microbiota-acting-on-human-proteases-involved-in-inflammatory-bowel-diseases
#18
Héla Mkaouar, Nizar Akermi, Vincent Mariaule, Samira Boudebbouze, Nadia Gaci, Florette Szukala, Nicolas Pons, Josan Marquez, Ali Gargouri, Emmanuelle Maguin, Moez Rhimi
BACKGROUND: In eukaryotes, the serpins constitute a wide family of protease inhibitors regulating many physiological pathways. Many reports stressed the key role of serpins in several human physiopathologies including mainly the inflammatory bowel diseases. In this context, eukaryotic serpins were largely studied and their use to limit inflammation was reported. In comparison to that, bacterial serpins and mainly those from human gut microbiota remain poorly studied. RESULTS: The two genes encoding for putative serpins from the human gut bacterium Eubacterium sireaum, display low sequence identities...
November 29, 2016: Microbial Cell Factories
https://www.readbyqxmd.com/read/27891183/complete-genome-of-vibrio-parahaemolyticus-forc014-isolated-from-the-toothfish
#19
Sojin Ahn, Han Young Chung, Sooyeon Lim, Kwondo Kim, Suyeon Kim, Eun Jung Na, Kelsey Caetano-Anolles, Ju-Hoon Lee, Sangryeol Ryu, Sang Ho Choi, Heebal Kim
BACKGROUND: Foodborne illness can occur due to various pathogenic bacteria such as Staphylococcus aureus, Escherichia coli and Vibrio parahaemolyticus, and can cause severe gastroenteritis symptoms. In this study, we completed the genome sequence of a foodborne pathogen V. parahaemolyticus FORC_014, which was isolated from suspected contaminated toothfish from South Korea. Additionally, we extended our knowledge of genomic characteristics of the FORC_014 strain through comparative analysis using the complete sequences of other V...
2016: Gut Pathogens
https://www.readbyqxmd.com/read/27888333/fusion-of-agarase-and-neoagarobiose-hydrolase-for-mono-sugar-production-from-agar
#20
Bassam Alkotaini, Nam Soo Han, Beom Soo Kim
In enzymatic saccharification of agar, endo- and exo-agarases together with neoagarobiose hydrolase (NABH) are important key enzymes for the sequential hydrolysis reactions. In this study, a bifunctional endo/exo-agarase was fused with NABH for production of mono-sugars (D-galactose and 3,6-anhydro-L-galactose) from agar using only one fusion enzyme. Two fusion enzymes with either bifunctional agarase (Sco3476) or NABH (Zg4663) at the N-terminus, Sco3476-Zg4663 (SZ) and Zg4663-Sco3476 (ZS), were constructed...
November 25, 2016: Applied Microbiology and Biotechnology
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